We first analyzed data on H3K4me3 modification from mouse testis and liver, generated from the Hop2-/ strain (Smagulova et al. 2011).
Administration of rapamycin or control vehicle began on postnatal day (P 12 and ended on P25, a period of mouse development in which the testis generates the first sets of meiotic and post-meiotic germ cells [ 29].
While pluripotent stem cells had been successfully generated previously from neonatal mouse testis, it remained unclear until recently whether the same could be achieved for wild-type adult SSCs in long-term culture [ 66].
For mouse testis and isolated germ cell types, microarray expression data, generated using the Affymetrix chip mouse genome 430 2.0 [18], were obtained from NCBI GEO (http://www.ncbi.nlm.nih.gov/geo/) using GEO accession number GSE4193.
To generate pEGFP-C1-SNX5, the full length cDNA of SNX5 was amplified from mouse testis cDNA and the resulting PCR product was cloned into KpnI/ BamHI sites of pEGFP-C1.
The full length cDNA of SNX1 was amplified from mouse testis cDNA and the resulting PCR product was cloned into EcoRI/ XmaI sites of pEGFP-C1 (Clontech, USA) to generate pEGFP-C1-SNX1 construct.
We also analyzed a genome-wide map of H3K4me3 and H3K27me3 from human sperm (Hammoud et al. 2009) and mouse testis (Cui et al. 2012), as well as H3K4me3 modification map from mouse testis and liver (Smagulova et al. 2011) generated by chromatin-immunoprecipitation sequencing methods.
We measured DNA double-strand breaks (DNA DSBs) and oxidative stress parameters including malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, and testis weight and sperm count at 12 h, 21 d and 35 d after irradiation in mouse testis.
To evaluate the system, 55 cDNA clones with predominantly unknown function were selected from a mouse testis cDNA-library.
A Sertoli cell line from mouse testis (42GPA9) was analyzed for cytoskeletal and Sex Hormone Binding Globilin (SHBG) changes by immunohistochemistry, for antioxidant content by RT-PCR and for culture medium lactate concentrations by protein chemistry.
